Zika Virus Research
RNAscope™ Applications
Zika Virus Research
As a result of the Zika virus outbreak, researchers are accelerating research into the biology of the virus and the pathology of human infection with the goals of understanding the causes of the clinical syndrome, developing reliable detection methods and finding effective treatments.
Why use RNAscope™ assay for zika virus research?
RNAscope™ ISH, an innovated ISH assay, offers researchers a robust and accurate method to detect even low levels of viral RNA in virtually any tissue samples. The ability to detect the virus within the context of the infected tissue will advance the understanding of the zika viral biology.
Morphologic context - Performed in situ, RNAscope™ assay preserves the morphology of the infected tissues, enabling detection of the viral RNA in the morphological context. Miner et al., 2016 demonstrated this in detection of Zika in eyes of mouse fetuses
Proven for viral detection - Numerous publications citing the use of RNAscope™ in situ hybridization in viral studies spanning HPV, EPV, HIV, SIV, HCV, MERS etc. RNAscope™ assay enables virologists to bypass time consuming and often non-yielding antibody development.
Proven for viral replication detection and localization - read Bhatnagar J et al 2017
Versatile probe design for low viral count - When working with zika or other viruses of extremely low copy number, ACD can customize the probe design strategy to offset this challenge; for example by designing probes with more than the typical 20 Z probe pairs.
Specificity - The highly specific, proprietary RNAscope™ double Z probe design, eliminates cross reactivity with other flavivirus RNA, enabling reliable differentiation from highly homologous viruses such as dengue.
Sensitivity - The ultrahigh sensitivity of RNAscope™ assay and ability to detect single RNA molecules makes it ideal for in situ detection of low level tissue infection. Read Rosenberg et al (2017)
Featured Publications
A number of key customers have used the RNAscope™ ISH Assay in their published research findings.
| Date/Journal | Title and details |
|---|---|
July 2016
| New England Journal of Medicine Miscarriage associated with Zika virus infection van der Eijk AA et al detected ZIKA viral RNA with the use of virus-specific RNAscope™ probes that revealed staining in the amniotic epithelium and the fetal mesenchymal cells, particularly in the perichondrium. |
SEPT 2016
| Cell Reports Zika virus infection in mice causes panuveitis with shedding of virus in tears Miner et al., 2016 detected Zika virus in eyes of mouse fetuses - specifically in the retina, optic nerve and cornea (figure 6). |
SEPT 2016 | Archives of Path & Lab Med Rosenberg et al (2017) applied in situ hybridization using a Zika virus RNAscope™ Probe 463781 demonstrated scattered, strongly positive staining cells within the villous stroma of the chorionic villi (Figure 4), which were presumably Hofbauer cells. |
OCT 2016
| Nature Medicine Zika viral dynamics and shedding in rhesus and cynomolgus macaques Osuna et al used RNAscope™ ISH assay to study ZIKV dynamics and tropism in tissues. Multiple anatomic tissues from each animal were prepared and analyzed by RNAscope, including lymph nodes (LNs) (inguinal, axillary, mesenteric), male genital tract (seminal vesicles, testes, prostate), female genital tract (uterus, ovaries), gastrointestinal tract (jejunum, colon), liver, kidney, bladder, lung, bone marrow and CNS (parietal lobe, basal nuclei, hippocampus, cerebellum) figure 4. The RNAscope probes with 87 Z pairs were designed against a consensus sequence comprise of 4 different genomes. |
OCT 2016
| Nature Zika virus infection damages the testes in mice Govero et al (2016) used RNAscope™ Probe Catalog #467871 to determine Zika viral tropism. RNAscope in situ hybridization (ISH) was applied to detect viral RNA at in cells of the testis at day 7 after infection (Fig 1), day 14 (Fig 2), day 21 (Fig 3) and day 28 (Fig 4). Cells of the testis include mature lumenal sperm, and on cilia layering the inner lumen of the epididymis. |
NOV 2016
| Nature Neutralizing human antibodies prevent Zika virus replication and fetal disease in mice. Sapparapu et al (2016) used RNAscope Probe 467771
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DEC 2016
| Emerging Infectious Disease Zika virus RNA replication and persistence in brain and placental tissue Bhatnagar J et al (2017) used RNAscope Probes 468361 and 469541 (designed against the sense and anti-sense strands) and demonstrated not only detection of the virus but also detection of viral replication. They isolated the viral replication to neuronal cells in fetal brain tissues. |
JAN 2017
| PLOS Smith (2017) and a team of researchers at the United States Army Medical Research Institute of Infectious Diseases applied RNAscope 2.5 HD RED to visualize Zika Virus in mouse models. Their goal is to development of well-characterized animal models that recapitulate human disease. RNAscope ISH data was presented throughout the paper in Fig 3 (ZIKV in Cerebrum) . Fig 4 (ZIKV in hippocampus), Fig 5 (ZIKV in spinal cord), Fig 7 (ZIKV in spleen). Using this model, they provide a detailed description of the ZIKV-associated pathologic changes, which mirrors the neuropathogenic properties of ZIKV in humans. |
ZIKA Virus Genome Structure
The zika virus is a sense single-stranded RNA virus (sssRNA) (also described as SSRNA positive sense virus, no DNA stage). Its >10,000 bases code for three structural proteins (capsid (C), precursor membrane (prM), envelope (E) and seven nonstructural proteins (NS). The NS2A, NS2B, NS4A, and NS4B proteins are smaller, hydrophobic proteins while NS1, NS3, and NS5 are large and highly conserved.
ACD can design RNAscope™ ISH probes against unique regions of these publically available sequences or proprietary sequences that you may have. We can design it to the sense or antii-sense strand. Depending on your study, the probe design can target specific gene regions or span across the genome. We will also work with you to design positive and negative controls.
